Our strategy has been to identify FDA-approved drugs or investigational new drugs (IND) that might be used in off-label applications to treat viral encephalitis so that the pathway to human treatment would be shorter. Preliminary data suggests that glutamate receptor antagonists, modulators of nitric oxide, or viralreactive antibodies might be used to treat viral encephalitis after the virus has entered the brain. The objective of this project is to identify biochemical, viral or cellular markers in the cerebrospinal fluid (CSF) or serum for benchmark pathological events in live animals that are needed for a) effectively treating viral encephalitis, b) determining the order and timing of these pathological events, c) clinically managing the treatment, and d) planning clinical trials. Additionally, the objective is to use these markers to appropriately treat West Nile virus (WNV) encephalitis and Western equine encephalitis (WEE) using FDA-approved drugs or IND identified in our current screening NIH contracts at USD. Specific Aim #1: Identify and correlate the time-course of viral expression, blood-brain-barrier (BBB) permeabilization, necrosis, neuron apoptosis and neuro-inflammation in specific portions of the brain and at different stages of disease. Markers for disease outcome and therapy in CSF and serum have not been adequately identified. Specific Aim #2: Compare CSF and serum 2D-gel electrophoresis proteins and classical clinical markers with disease parameters at different stages. Based on our preliminary results, proteins will be identified in CSF and serum at different stages of disease from uninfected and WNV- and WEE-infected hamsters using 2-D electrophoresis fluorescent-labeled protein technology (DICE). Disease protein-markers will be identified by mass spectrophotometry coupled with proteome bioinformatics. Specific Aim #3: Use CSF or serum markers to identify, and most effectively administer, therapies for WNV or WEE disease. Therapeutics will be evaluated in the hamster model at different stages of disease using clinical or CSF/serum markers and will be administered in the brain by convection-enhanced delivery (CED). FDA-approved or IND candidates showing efficacy will be communicated with University of Utah Infectious Disease Unit for possible off-label administration to WNV-infected patients or to companies with compatible technologies. Project interactions: This and other projects within the USU program will collaborate with the Infectious Disease unit at the U of U, with treatment of viral encephalitis, with aerolization of drugs and virus at the Animal Models Core, and with industry, i.e., IVAX Research and Axonix USA.